Department of Chemical Engineering, Ferdowsi University of Mashhad (FUM), Mashhad, Iran.
Department of Biology, Ferdowsi University of Mashhad (FUM), Mashhad, Iran.
J Tissue Eng Regen Med. 2019 Oct;13(10):1861-1871. doi: 10.1002/term.2936. Epub 2019 Aug 27.
Previous researches have emphasized on suitability of decellularized tissues for regenerative applications. The decellularization of cartilage tissue has always been a challenge as the final product must be balanced in both immunogenic residue and mechanical properties. This study was designed to compare and optimize the efficacy of the most common chemical decellularization treatments on articular cartilage. Freeze/thaw cycles, trypsin, ethylenediaminetetraacetic acid (EDTA), sodium dodecyl sulfate (SDS), and Triton-X 100 were used at various concentrations and time durations for decellularization of bovine distal femoral joint cartilage samples. Histological staining, scanning electron microscopy, DNA quantification, compressive strength test, and Fourier-transform infrared spectroscopy were performed for evaluation of the decellularized cartilage samples. Treatment with 0.05% trypsin/EDTA for 1 day followed by 3% SDS for 2 days and 3% Triton X-100 for another 2 days resulted in significant reduction in DNA content and simultaneous maintenance of mechanical properties. Seeding the human adipose-derived stem cells onto the decellularized cartilage confirmed its biocompatibility. According to our findings, an optimized physiochemical decellularization method can yield in a nonimmunogenic biomechanically compatible decellularized tissue for cartilage regeneration application.
先前的研究强调了脱细胞组织在再生应用中的适宜性。软骨组织的脱细胞化一直是一个挑战，因为最终产物在免疫原性残留和机械性能方面必须保持平衡。本研究旨在比较和优化最常见的化学脱细胞处理方法对关节软骨的效果。采用冻融循环、胰蛋白酶、乙二胺四乙酸（EDTA）、十二烷基硫酸钠（SDS）和 Triton-X100 分别在不同浓度和时间下处理牛远端股骨关节软骨样本以进行脱细胞化。通过组织学染色、扫描电子显微镜、DNA 定量、压缩强度测试和傅里叶变换红外光谱对脱细胞软骨样本进行评估。结果表明，用 0.05%胰蛋白酶/EDTA 处理 1 天，然后用 3% SDS 处理 2 天，再用 3% Triton X-100 处理 2 天，可显著降低 DNA 含量，同时保持机械性能。将人脂肪源性干细胞接种到脱细胞软骨上证实了其生物相容性。根据我们的发现，优化的物理化学脱细胞方法可以产生一种非免疫原性、生物力学兼容的脱细胞组织，用于软骨再生应用。
